Next Workshop on Epigenetic Profiling ChIP & MCIp

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1 Next Workshop on Epigenetic Profiling ChIP & MCIp 18-21st October 2010 Organized by the Division of Epigenomics and Cancer Risk Factors DKFZ Heidelberg, and Diagenode s training team Have you heard about these exciting techniques? Now is your chance to learn how to perform these protocols and bring the technology back to your lab! Limited number of places! 1

2 WORKSHOP DETAILS How it works in brief Methyl CpG immunoprecipitation (MCIp) is performed as follows: Genomic DNA from cultured cells or tissues is prepared and fragmented by sonication. Chromatin IP is performed as follows: Chromatin is first treated with formaldehyde to fix DNA-bound proteins to the DNA. The chromatin is subsequently sheared into small fragment sizes (200bp 1kb). Genomic DNA fragmentation (Bioruptor ) ro ein cross in ing e sis onica ion or en me diges ion methyldna capture, robot (SX-8G IP- Star ) or lab bench; fractionation by degree of methylation ragmen ed c roma in mm no reci i a ion i s ecific an i od mm ne reci i a e ma eria rifica ion na sis of o nd methyldna quantitation by qpcr on imprinted genes icroarra e encing Using the IP-Star robot or performed on lab bench, fragmented DNA is captured by MBD2 protein coupled to magnetic beads. Coupled DNA is fractionated according to degree of methylation using an increasing salt gradient. Following DNA purification, fractions are ready for any subsequent analysis (e.g. qpcr, microarray or next generation sequencing). Immunoprecipitation follows using a specific ChIP-grade antibody. Crosslinking is reversed and the DNA can be purified using different approaches (e.g. magnetic bead IPure kit). The purified DNA is analyzed to identify the genomic regions where the specific protein is located using, for example, next generation sequencing (ChIPseq), qpcr (ChIP) or hybridization on a microarray (ChIPchip). During the workshop, we ll go through each step of both techniques and talk about optimization and potential pitfalls. You ll be provided with everything you need at the workshop to carry through an entire chromatin and methylated DNA immunoprecipitation experiment in a model system. All you need to bring is a list of questions and your enthusiasm to learn new technologies! To enhance our workshop we have invited 3 scientists working in the field of epigenomics to present their data and discuss their experiences in epigenetic analyses such as MCIp and MeDIP followed by microarray analyses or next generation sequencing. You will also enjoy the friendly and professional atmosphere created by DIAGENODE s team during and after sessions Training, reagents and registration is included in the price (accommodation and travel must be booked separately; see registration form page 5). 2

3 Monday 18th October h15 9h30 / 10h30 10h45 / 12h00 13h30 / 15h30 Welcome "Introduction: Epigenetic chromatin modifications" "Presentation of Practical Training" Workshop ChIP: chromatin crosslinking Workshop MCIp: DNA sonication; gel analysis of DNA 15h30 / 16h30 Conference 1 16h45 / 18h00 Workshop ChIP: chromatin sonication, o/n proteinase K digestion for quality control (QC) Workshop MCIp: MBD2-bead coupling Tuesday 19th October h00 / 12h00 13h30 / 15h30 Workshop ChIP: QC continued with phenol/chloroform extraction; gel analysis of DNA Workshop MCIp: DNA capture by MBD2, robot and lab bench Workshop ChIP: Immunoprecipitation Workshop MCIp: DNA purification 15h45/ 16h45 Conference 2 Wednesday 20th October h00 / 12h00 13h30 / 16h00 16h15 / 17h30 Workshop ChIP: DNA purification Workshop MCIp: qpcr Workshop ChIP: DNA purification continued Workshop ChIP: qpcr Workshop MCIp: analysis of qpcr results Thursday 21th October h00 / 10h00 Conference 3 10h15 / 11h30 11h45 / 12h45 Workshop ChIP: analysis of qpcr results Workshop MCIp: analysis of qpcr results continued Conclusions and discussion 3

4 Registration form ChIP MCIp Workshop October 18-21st 2010 Registration deadline: September 15th 2010 Registration fees: (accommodation not included) - Public or academic: 750 excl taxes - Private: 1,150 excl taxes The number of places is limited to 10 attendees per workshop! First Name: Last Name: Researcher Group leader Engineer Lecturer Medical Dr Technician PhD Student Post-doc Graduate student Other (precise) Academic sector Private sector Tel: Fax: Address: Name of the laboratory: Institution or Company: Invoicing address: Contact person: VAT number Fiscal / Tax id: Director s signature Applicant s signature 4

5 Form to be returned to: Fax: +49 (0) To allow us to understand and design the workshop to suit your needs, please fill in the following questionnaire: How did you hear about this Diagenode s workshops? Revue Web Site through a colleague Other, precise: Can you tell us more about your research project, ongoing or planned? If so, briefly describe it: Do you have specific needs or expectation? If so, briefly describe it: 5